title: PNA-NLS conjugates as single-molecular activators of target sites in double-stranded DNA for site-selective scission.
abstract: Artificial DNA cutters have been developed by us in our previous studies by combining two strands of pseudo-complementary peptide nucleic acid (pcPNA) with Ce(IV)-EDTA-promoted hydrolysis. The pcPNAs have two modified nucleobases (2,6-diaminopurine and 2-thiouracil) instead of conventional A and T, and can invade double-stranded DNA to activate the target site for the scission. This system has been applied to site-selective scissions of plasmid, λ-phage, E. coli genomic DNA, and human genomic DNA. Here, we have reported a still simpler and more convenient DNA cutter obtained by conjugating peptide nucleic acid (PNA) with a nuclear localization signal (NLS) peptide. This new DNA cutter requires only one PNA strand (instead of two) bearing conventional (non-pseudo-complementary) nucleobases. This PNA-NLS conjugate effectively activated the target site in double-stranded DNA and induced site-selective scission by Ce(IV)-EDTA. The complex formation between the conjugate and DNA was concretely evidenced by spectroscopic results based on time-resolved fluorescence. The target scission site of this new system was straightforwardly determined by the Watson-Crick base pairing rule, and mismatched sequences were clearly discriminated. Importantly, even highly GC-rich regions, which are difficult to be targeted by a previous strategy using pcPNA, were successfully targeted. All these features of the present DNA cutter make it promising for various future applications.
authors.affiliation: Life Science Center of Tsukuba Advanced Research Alliance, University of Tsukuba, 1-1-1 Tennoudai, Tsukuba, Ibaraki 305-8577, Japan.
authors.forename: Yuichiro
authors.initials: Y
authors.lastname: Aiba
authors.forename: Yuya
authors.initials: Y
authors.lastname: Hamano
authors.forename: Wataru
authors.initials: W
authors.lastname: Kameshima
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Araki
authors.forename: Takehiko
authors.initials: T
authors.lastname: Wada
authors.forename: Alessandro
authors.initials: A
authors.lastname: Accetta
authors.forename: Stefano
authors.initials: S
authors.lastname: Sforza
authors.forename: Roberto
authors.initials: R
authors.lastname: Corradini
authors.forename: Rosangela
authors.initials: R
authors.lastname: Marchelli
authors.forename: Makoto
authors.initials: M
authors.lastname: Komiyama
mesh_terms: D020137:Base Pair Mismatch; D001483:Base Sequence; D002563:Cerium / Q000737:chemistry; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D004492:Edetic Acid / Q000737:chemistry; D006801:Humans; D019913:Nuclear Localization Signals; D020135:Peptide Nucleic Acids / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
chemical_list: D019913:Nuclear Localization Signals; D020135:Peptide Nucleic Acids; D002563:Cerium; D004247:DNA; D004492:Edetic Acid